Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Mutant p53 antibody [E47] - BSA and Azide free (ab247264)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E47] to Mutant p53 - BSA and Azide free
  • Suitable for: ICC, WB, Flow Cyt, IP, IHC-P
  • Reacts with: Human

Overview

  • Product name

    Anti-Mutant p53 antibody [E47] - BSA and Azide free
    See all Mutant p53 primary antibodies

  • Description

    Rabbit monoclonal [E47] to Mutant p53 - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    This antibody recognises human mutant forms of p53 but not human p53 wild type. It showed a negative signal on wildtype p53 cell lines (HepG2, A549, MCF-7) and a positive signal on mutant p53 cell lines (T47-D, Raji, A431) in WB.

  • Tested applications

    Suitable for: ICC, WB, Flow Cyt, IP, IHC-Pmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A431 cell lysate. ICC/IF: A431 cells. IHC-P: Human lung carcinoma tissue. Flow Cyt: HEK-293 cells IP: HEK-293 whole cell lysate.

  • General notes

    ab247264 is the carrier-free version of ab32509 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    Ab247264 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Images

  • Western blot - Anti-Mutant p53 antibody [E47] - BSA and Azide free (ab247264)
    Western blot - Anti-Mutant p53 antibody [E47] - BSA and Azide free (ab247264)
    All lanes : Anti-Mutant p53 antibody [E47] (ab32509) at 1/200 dilution

    Lane 1 : A549 (Human lung carcinoma epithelial cell), Whole cell lysate
    Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell), Whole cell lysate
    Lane 3 : MCF-7 (Human breast adenocarcinoma epithelial cell), Whole cell lysate
    Lane 4 : T-47D (Human ductal breast epithelial tumor epithelial cell), Whole cell lysate
    Lane 5 : A431 (Human epidermoid carcinoma epithelial cell), Whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 44 kDa



    This data was developed using ab32509, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 180 seconds

    Lane 1-3: Wildtype p53 cell lines

    Lane 4-5: Mutant p53 cell lines

    Observed MW: 50kDa, 39kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [E47] - BSA and Azide free (ab247264)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [E47] - BSA and Azide free (ab247264)
    This data was developed using ab32509, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded human lung carcinoma sections labeling mutant p53 with ab32509 at 1/2000 dilution (0.49 μg/mL). Sections were counterstained with Hematoxylin. Goat Anti-Rabbit IgG H&L (HRP Polymer) was used as the secondary antibody. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).
  • Immunocytochemistry - Anti-Mutant p53 antibody [E47] - BSA and Azide free (ab247264)
    Immunocytochemistry - Anti-Mutant p53 antibody [E47] - BSA and Azide free (ab247264)
    This data was developed using ab32509, the same antibody clone in a different buffer formulation.Immunocytochemistry/Immunofluorescence analysis of A431 (human epidermoid carcinoma) labelling mutant p53 with purified ab32509 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (ab150077). Nuclei counterstained with DAPI (blue). Control: PBS only.
  • Flow Cytometry - Anti-Mutant p53 antibody [E47] - BSA and Azide free (ab247264)
    Flow Cytometry - Anti-Mutant p53 antibody [E47] - BSA and Azide free (ab247264)

    This data was developed using ab32509, the same antibody clone in a different buffer formulation.
    Flow Cytometry analysis of HEK-293 (Human embryonic kidney epithelial cell) cells labeling Mutant p53 with purified ab32509 at 1/100 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cells without incubation with primary antibody and secondary antibody (Blue).

  • Immunoprecipitation - Anti-Mutant p53 antibody [E47] - BSA and Azide free (ab247264)
    Immunoprecipitation - Anti-Mutant p53 antibody [E47] - BSA and Azide free (ab247264)

    This data was developed using ab32509, the same antibody clone in a different buffer formulation.
    Purified ab32509 at 1/50 dilution (2µg) immunoprecipitating Mutant p53 in HEK-293 whole cell lysate.
    Lane 1 (input): HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate 10µg
    Lane 2 (+): ab32509 + HEK-293 whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32509 in HEK-293 whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
    Blocking Buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM/TBST.
    Observed band size: 53 kDa

  • Western blot - Anti-Mutant p53 antibody [E47] - BSA and Azide free (ab247264)
    Western blot - Anti-Mutant p53 antibody [E47] - BSA and Azide free (ab247264)
    Anti-Mutant p53 antibody [E47] (ab32509) at 1/1000 dilution + A431 lysate

    Predicted band size: 44 kDa
    Observed band size: 53 kDa
    why is the actual band size different from the predicted?



    This data was developed using ab32509, the same antibody clone in a different buffer formulation.

     

  • Anti-Mutant p53 antibody [E47] - BSA and Azide free (ab247264)
    Anti-Mutant p53 antibody [E47] - BSA and Azide free (ab247264)

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